title: Imaging energy status in live cells with a fluorescent biosensor of the intracellular ATP-to-ADP ratio.
abstract: The ATP:ADP ratio is a critical parameter of cellular energy status that regulates many metabolic activities. Here we report an optimized genetically encoded fluorescent biosensor, PercevalHR, that senses the ATP:ADP ratio. PercevalHR is tuned to the range of intracellular ATP:ADP expected in mammalian cells, and it can be used with one- or two-photon microscopy in live samples. We use PercevalHR to visualize activity-dependent changes in ATP:ADP when neurons are exposed to multiple stimuli, demonstrating that it is a sensitive reporter of physiological changes in energy consumption and production. We also use PercevalHR to visualize intracellular ATP:ADP while simultaneously recording currents from ATP-sensitive potassium (KATP) channels in single cells, showing that PercevalHR enables the study of coordinated variation in ATP:ADP and KATP channel open probability in intact cells. With its ability to monitor changes in cellular energetics within seconds, PercevalHR should be a versatile tool for metabolic research.
authors.affiliation: Department of Neurobiology, Harvard Medical School, 220 Longwood Avenue, Boston, Massachusetts 02115, USA.
authors.forename: Mathew
authors.initials: M
authors.lastname: Tantama
authors.forename: Juan Ramón
authors.initials: JR
authors.lastname: Martínez-François
authors.forename: Rebecca
authors.initials: R
authors.lastname: Mongeon
authors.forename: Gary
authors.initials: G
authors.lastname: Yellen
mesh_terms: D000244:Adenosine Diphosphate / Q000032:analysis* / Q000096:biosynthesis; D000255:Adenosine Triphosphate / Q000032:analysis* / Q000096:biosynthesis; D000818:Animals; D019843:Archaeal Proteins / Q000235:genetics / Q000378:metabolism; D001253:Astrocytes / Q000378:metabolism* / Q000648:ultrastructure; D015374:Biosensing Techniques; D002460:Cell Line; D004622:Embryo, Mammalian; D004734:Energy Metabolism / Q000502:physiology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D006801:Humans; D054086:KATP Channels / Q000235:genetics / Q000378:metabolism; D051379:Mice; D036641:Microscopy, Fluorescence, Multiphoton; D057054:Molecular Imaging; D009474:Neurons / Q000378:metabolism* / Q000648:ultrastructure; D061251:Primary Cell Culture; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D059010:Single-Cell Analysis
chemical_list: D019843:Archaeal Proteins; C449645:GlnK1 protein, Methanosarcina mazei; D054086:KATP Channels; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate